This Shared Instrument Grant proposal is for the purchase of an Amnis ImageStream instrument that combines the power of flow cytometry with the capture of high resolution digital images of cells in flow. The ImageStream is able to detect four colors of fluorescence as well as capture brightfield and darkfield imagery of each cell. The ImageStream does not replace any existing flow cytometric or imaging equipment, but rather, provides a complementary technology that will greatly facilitate the research of the grant participants in applications such as nuclear translocation studies in mixed populations of cells, apoptosis, cell cycle and mitosis studies, marker co-localization, intracellular trafficking, cell:cell interactions, fluorescent in situ hybridization and gene expression/co-localization. In addition to the instrument, a stabilizing table for the ImageStream is also requested. A Dell Workstation that provides powerful data management is requested, as is a multi-user software license for the IDEAS software. The new ImageStream will be housed in the Flow Cytometry Core Facility at the New Jersey Medical School and will be operated and maintained by an experienced flow cytometry technician Ms. Angelika Batista. The Principal Investigator, Dr. Patricia Fitzgerald-Bocarsly will provide administrative and scientific oversight in consultation with a Faculty Advisory Committee. The ImageStream will be primarily utilized by the applicant group of thirteen highly-productive scientists from diverse departments and biomedical science disciplines at the NJMS. They include: 1) Azzam, Edouard - Damage signaling from irradiated to non-irradiated cells; 2) Fitzgerald-Bocarsly, Patricia - Plasmacytoid dendritic cells in HIV pathogenesis; 3) Gause, William - Cytokine gene expression during in vivo immune response; 4) Howell, Roger - Effects of nonuniform distributions of radioactivity; 5) Kotenko, Sergei - Role of interferon lambdas in antiviral response; 6) Lambert, Muriel - DNA repair defect in Fanconi Anemia Group A; 7) Leibovich, Joseph - Toll-like receptors, adenosine and angiogenesis; 8) Lukac, David - DNA binding mechanism of the KSHV lytic switch protein and kinetics of KSHV gene expression; 9) Rameschwar, Pranela - Vaccine hurdle to anthrax and the emerging immune system; IFN-gamma in the neuroimmune-mesenchymal stem cell axis; and Breast cancer-bone marrow stromal interactions; 10) Rohowsky-Kochan, Christine - Cytokine secretion profile of autoreactive T cells; 11) Salgame, Padmini - Induction of Th1 immunity in tuberculosis; 12) Studzinski, George - Vitamin D analogs as adjuvants in chemotherapy of cancer; and 13) Molina, Carlos - Ras-Mediated proteasomal degradation of ICER in melanomas. This group of investigators is anticipated to utilize almost 100% of the total usage of the instrument. Remaining instrument time will be allocated to NIH-funded investigators or other individuals at the Newark campus. Instrument time may be made available to non-UMDNJ investigators from time-to-time when the ImageStream is not otherwise being used. [unreadable] [unreadable] [unreadable]